NIGMS Equipment Supplement on R01 GM098885

R01 GM098885 的 NIGMS 设备补充

基本信息

批准号：
10794115
负责人：
James L Keck
金额：
$ 1.88万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-30 至 2027-06-30
项目状态：
未结题

项目摘要

Abstract: DNA replication restart pathways reload cellular DNA replication complexes onto replication forks that have been prematurely terminated, forming an essential link between DNA repair and replication. The proteins that drive these reactions, referred to as the primosome or the Replication Restart Proteins, recognize the structures of abandoned replication forks and reload the DNA replication machinery specifically at these sites. The replication restart process is regulated to ensure loading fidelity and to avoid over-replication that could arise from initiating replication at improper DNA structures. In spite of the broad biological importance of this process, the mechanisms underlying DNA replication restart and its regulation remain poorly understood. Our proposal combines structural, biochemical, and genetic approaches to define the structural and cellular mechanisms of DNA replication restart. Our overall objective is to determine the mechanisms that govern each step of the process, from recognition of diverse structures of abandoned DNA replication forks, to primosome assembly, and finally to reloading the DNA replication machinery. Aim 1 will define the structures of primosome complexes with replication forks, which will provide new insights into structure-specific recognition of DNA replication forks and the mechanisms that regulate DNA replication restart. Structural advances are coupled to genetic studies that will define the cellular mechanisms of action of primosome proteins in bacteria. Aim 2 blends structural and genetic studies to focus on maturation of primosome complexes and their activities in reloading the replicative helicase back onto replication forks. Completion of our specific aims will define the molecular and cellular mechanisms that mediate and regulate bacterial DNA replication restart.

抽象的： DNA复制重新启动途径将细胞DNA复制复合复合复合复合物重新升级到已过早终止的复制叉，形成了基本链接在DNA修复和复制之间。驱动这些反应的蛋白质，称为作为原始体或复制重新启动蛋白质，识别的结构放弃的复制叉，并重新加载DNA复制机械专门这些站点。调节复制重新启动过程以确保加载保真度和避免在不适当的DNA上启动复制而产生的过度复制结构。尽管这一过程具有广泛的生物学重要性，但机制基本的DNA复制重新启动及其调控仍然很少理解。我们的建议结合结构，生化和遗传方法来定义结构 DNA复制的细胞机制重新启动。我们的总体目标是确定从认识到多样化的过程的每个步骤的机制废弃的DNA复制叉的结构，将原始体组装到最终重新加载DNA复制机械。 AIM 1将定义原始体的结构具有复制叉的复合物，这将为特定于结构的新见解提供新的见解识别DNA复制叉和调节DNA复制的机制重新启动。结构的进步与遗传研究结合，该研究将定义细胞原始蛋白在细菌中的作用机理。 AIM 2融合结构和遗传研究专注于原始体复合物的成熟及其在将复制性解旋酶重新加载回复制叉。完成我们的特定目标将定义介导和调节的分子和细胞机制细菌DNA复制重新启动。

项目成果

期刊论文数量（29）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The molecular coupling between substrate recognition and ATP turnover in a AAA+ hexameric helicase loader.

DOI：
10.7554/elife.64232
发表时间：
2021-05-26
期刊：
eLife
影响因子：
7.7
作者：
Puri N;Fernandez AJ;O'Shea Murray VL;McMillan S;Keck JL;Berger JM
通讯作者：
Berger JM

A guanine-flipping and sequestration mechanism for G-quadruplex unwinding by RecQ helicases.

DOI：
10.1038/s41467-018-06751-8
发表时间：
2018-10-10
期刊：
Nature communications
影响因子：
16.6
作者：
Voter AF;Qiu Y;Tippana R;Myong S;Keck JL
通讯作者：
Keck JL

Interaction with single-stranded DNA-binding protein localizes ribonuclease HI to DNA replication forks and facilitates R-loop removal.

DOI：
10.1111/mmi.14529
发表时间：
2020-09
期刊：
Molecular microbiology
影响因子：
3.6
作者：
Wolak C;Ma HJ;Soubry N;Sandler SJ;Reyes-Lamothe R;Keck JL
通讯作者：
Keck JL

Structural and Functional Studies of H. seropedicae RecA Protein - Insights into the Polymerization of RecA Protein as Nucleoprotein Filament.

DOI：
10.1371/journal.pone.0159871
发表时间：
2016
期刊：
PloS one
影响因子：
3.7
作者：
Leite WC;Galvão CW;Saab SC;Iulek J;Etto RM;Steffens MB;Chitteni-Pattu S;Stanage T;Keck JL;Cox MM
通讯作者：
Cox MM

Positive Charges Are Important for the SOS Constitutive Phenotype in recA730 and recA1202 Mutants of Escherichia coli K-12.

正电荷对于大肠杆菌 K-12 的recA730 和recA1202 突变体中的SOS 组成表型很重要。